基质辅助激光解吸电离飞行时间串联质谱法分析人乳β-酪蛋白新生儿体外消化模型多肽组

通过体外模拟新生儿消化道条件,用基质辅助激光解吸离子化飞行时间串联质谱法(MALDI-TOF/TOF)探究人乳β-酪蛋白消化后的多肽组.在离子源加速电压为20 kV,激光波长337 nm,激光频率200 Hz,离子延迟提取时间330 ns,质谱信号单次扫描累加2000次条件下,扫描质量范围m/z 500 ～ 5000的肽段.结果表明,消化后得到26个肽段,分子量集中在1000～4000Da.与已知功能的肽段序列进行对比,人乳β-酪蛋白在新生儿体外模型消化后未产生与已知功能的活性肽序列匹配的肽段,但发现9个肽前体,其中含4个血管紧张素转化酶(ACE)抑制肽前体,2个酪蛋白磷酸肽(CPP)前体,2个抗氧化肽前体,1个免疫活性肽前体,由酶切位点推测分析,肽前体可以继续在蛋白酶作用下转化为生物活性肽.     

The synergistic chaperoning operation in a Bi-chaperone system consisting of alpha-crystallin and beta-casein: bovine pancreatic insulin as the target protein

While chaperone activity of alpha-crystallin (α-Crs) is important in maintaining lens transparency that of beta-casein (β-CN) is vital to prevent the development of corpora amylacea (accumulation of amyloid deposits in mammary glands). These two chaperone proteins are amphiphilic, each contains distinct polar and non-polar regions in the structure. While polar domain of α-Crs is highly electropositive, the counterpart domain in β-CN is strongly electronegative. In this study a Bi-chaperone system consisting of α-Crs and β-CN with different molar ratios were used to prevent the chemical-induced insulin aggregation spectroscopically. As shown, α-Crs and β-CN in the Bi-chaperone system exhibit synergistic chaperoning operation which strongly depends to the specific ratio of the chaperone components. The results of both fluorescence study and native gel electrophoresis confirmed the non-covalent interactions between α-Crs and β-CN. Consequently the synergistic activity can be explained with the possible electrostatic interactions between their polar/charged domains which bring them in close proximity, allowing their synergistic chaperoning operation in the Bi-chaperone system.